Chapter title |
Cell In Situ Zymography: Imaging Enzyme–Substrate Interactions
|
---|---|
Chapter number | 12 |
Book title |
Zymography
|
Published in |
Methods in molecular biology, June 2017
|
DOI | 10.1007/978-1-4939-7111-4_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7109-1, 978-1-4939-7111-4
|
Authors |
Aastha Chhabra, Vibha Rani |
Editors |
Jeff Wilkesman, Liliana Kurz |
Abstract |
Zymography has long been used for the detection of substrate-specific enzyme activity. In situ zymography (ISZ), an adaptation from the conventional substrate zymography, is a widely employed technique useful for the detection, localization, and estimation of enzyme-substrate interactions in tissues. Here, we describe a protocol to detect 'in position' matrix metalloproteinase (MMP) activity in cells utilizing H9c2 cardiomyoblasts as a model. This technique is primarily adopted from the method used for histological sections and is termed as 'Cell in situ Zymography'. It is a simple, sensitive, and quantifiable methodology to assess the functional activity of an enzyme 'on site/in position' in cell culture. |
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Demographic breakdown
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Unknown | 1 | 13% |
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